Tag Archives: MRC Centre for Developmental Neurobiology

Once I arrived at Kings College London, I was greeted by Dr Laura Andreae. We went up to her office and she talked me through her journey into science, specifically research. Surprisingly she wasn’t always set on doing research as prior to her current positions, she was a medic, having studied medicine at Cambridge University. She then did a Masters and fell in love with research, leading her eventually to Kings College London, where she lectures and supervises PhD students. After introductions with the rest of the members of her team, I got to put on my own lab coat.

I got to work with a Master student, Pavylios, and we used different enzymes such as TbrIII to cut out DNA and place them back into plasmids. I got to see and use equipment I haven’t seen before and I was particularly surprised as to how small everything was, since they used such small scales from a little as 1 µl (microliter). We then left them for a couple of hours so the enzymes could do their job. He then gave me a rough overview of genetics so I could really understand how the process works.

What I really found cool was what Adam, a PhD student, was doing. He was looking at brain tissue under a microscope but to get a more clearer and detailed image, he uses thicker samples. The problem with this is that the thicker the sample, the less detail you are able to see, and so to overcome this he uses a method called CLARITY, which makes the brain tissue transparent. This was really cool and when it was suspended in glycerol, you would think that there was nothing in there!

I then had the opportunity to see some live cell imaging, looking at one of the transparent tissues under a very expensive microscope. Under the microscope I saw some very detailed pictures of neurons and some pretty pictures of blood vessels. I also got to see a microscope which was currently in the process of being built by Adam and his colleagues. It was nice to see the physics and engineering behind microscopes. I then got to work with Sam, who was another PhD student, and I got to see him plate glia cells which can take up to weeks to grow and culture, and the got to see these under the microscope.

After that, I then got to attend a lab meeting where one researcher presented her work and current findings so her colleagues could discuss her work and advise her on what to do next. It was nice to see that the researchers do in fact work together despite each of them having their own different and individual projects.

Then I got to go back to the lab and work on the DNA we had begun working with on Monday. We needed to purify the DNA so we added some chemicals and used the centrifuge to separate the proteins from the DNA. Once we had the DNA in a separate tube, we moved over to the next lab so we could use a photometer. A photometer allows us to measure a concentration for the DNA. Pavylios was happy with the results and concluded that it was a successful experiment. I also had the opportunity to stain some tissues. They were stained with a marker that shows up as fluorescent green under special microscopes. Then, we did some more centrifugation, where we had to make RNA from the DNA we obtained and so added several chemicals and used the centrifuge to separate the DNA from RNA. We then left this for a couple of days so we could analyse them under a microscope.

I thought this was a great opportunity to gain an insight into what STEM career entails and look forward to being able to do the same in the future. I would recommend in2science to any student that hopes to have a successful career in science.

During my research placement at the Medical Research Centre for Developmental Neurobiology with Dr Tara Keck, I was able to take part in various experiments and techniques from electrophysiology to measure the potential difference and current of neurons, using computer software ‘Fiji’ to analyse neurons that are Green Fluorescent Protein (GFP) positive in inhibitory cells and immunohistochemistry to identify specific proteins.

My last two days involved carrying out immunohistochemistry to detect Glial Fibrillary Acidic Protein (GFAP). This protein is usually used as a cell marker and is expressed in one of the two types of brain cells called neuroglia, particularly astrocytes.

The image below displays astrocytes that express GFAP in the subventricular zone. This region of the brain is recognized as being the site of neurogenesis, the growth of new neurons which can also be seen in the image as the collection of cells on the left hand side of the subventricular zone.

My two week experience led me to the discovery of what to expect in neuroscience research and the rewarding feeling of working alongside post doctors and PhD students. The work can become somewhat frustrating when the day does not go as planned however the images and results produced are very pleasing.

The research my laboratory focused on was finding important proteins or eliminating genes that could affect neurodegeneration. One particular disease which is focused on is dentatorubral-pallidoluysian atrophy (DRPLA). The mating scheme diagram you can see in the image below is a hypothetical one created as a practice for designing ones in the lab. My supervisor Catarina regularly designed the mating schemes for flies so that she would be able to generate flies with certain genotypes to observe. It was amazing to find that some of the diagrams she had to make involved crossing the different stocks of flies 11 times, just to get the desired genotype.

Morgan describes her time in the MRC Centre for Developmental Neurobiology at King’s College London.

Things were more than busy on my first Wednesday whilst working at the lab for my work placement at the MRC Centre for Developmental Neurobiology at King’s College, not only was it my turn to work my way through the protocol for preparing coffee for the department that morning, but it was dissection day for Ivana (my postdoctoral supervisor). That meant that I had the pleasure in assisting her with the oculomotor nerve dissection she had planned for that day. As part of her project she needed to isolate cranial motor neurons from the this nerve found in the brain and later grow these cells in a culture to observe how their axons are guided to their targets during development.

Thursday proved to be busier in the lab as first thing that morning we got stuck into electroporation. Electroporation is a method that uses an externally applied electrical field on the cells of interest to increase the electrical conductivity and permeability of the cell plasma membrane. It is a way of introducing a substance into a cell, here we used it to insert DNA into our chosen cell (a process known as transfection).

The second week of my neuroscience placement has seen me come into contact with some pioneering methods used within genetic science and many of the biosciences. One of which is the use of the polymerase chain reaction (PCR). This is a form of genotyping which allowed us to determine the differences in genotype of the individual organisms we were studying by examining their DNA sequences and comparing them. It involves artificial DNA replication so that samples of DNA can be amplified to generate millions of copies for genetic profiling.

Gel electrophoresis. Now this is cool stuff. This particular technique is a modern molecular biology method used to separate out DNA fragments based on their size so that these fragments can be used for identification and analysis – this can tell us which genes are present in a sample and whether the individual expresses this gene or not. It uses an agarose gel with wells cut into it to place the DNA samples into, this is covered in a buffer solution in an electrophoresis tank with electrodes attached at each end so that a current is passed through attracting the negatively charged DNA and separating it through the gel by base length.